Processes for the preparation of spectinomycin analogs, novel products and intermediates therein

ABSTRACT

The present invention relates to novel methods of preparing a wide variety of spectinomycin analogs and biologically acceptable salts thereof. Further, the invention relates both to novel intermediates and novel products therein. The novel products are spectinomycin analogs which can be used for the same biological purposes as spectinomycin. The processes of the invention provide for novel intermediates that are versatile and highly reactive exocyclic enones.

This application is a division of application Ser. No. 381,113, now U.S.Pat. No. 4,467,103, filed May 24, 1982.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel methods of preparing a widevariety of spectinomycin analogs and biologically acceptable saltsthereof. Further, the invention relates both to novel intermediates andnovel products therein. The novel products are spectinomycin analogswhich can be used for the same biological purposes as spectinomycin. Theprocesses of the invention provide for novel intermediates that areversatile and highly reactive exocyclic enones having formula I

wherein R₁ and R₂ are blocking groups, and

R is selected from the group consisting of hydrogen, alkyl of C₁ to C₂₀,inclusive, lower alkenyl, lower haloalkyl, lower aminoalkyl, loweralkynyl, and --(CH₂)_(n) --OX

wherein X is selected from the group consisting of hydrogen, loweralkyl, lower alkenyl, benzyl, and acyl;

n is an integer of from zero to four with the proviso that when n iszero --OX cannot be hydroxy.

Two methods are provided for the synthesis of the enones I byfunctionalization of the starting materials, i.e., protectedspectinomycin and protected spectinomycin analogs having formula III'.Such functionalization provides for Mannich bases of formula III" either(1) directly or (2) through novel intermediate enamines having formulaII. The Mannich bases give rise to the enones I. Finally, either theenones I or the enamines II are reacted with nucleophiles to obtain thewide variety of spectinomycin analogs including the novel products ofthe present invention.

R₁, R₂, and R in formulae I, II, III', and III" are as described above.

2. Description of the Art

Spectinomycin is a known antibiotic having the formula III. Amicrobiological preparation may be found in U.S. Pat. No. 3,234,092. Achemical synthesis including the nitrogen protected form, now among thestarting materials of the present invention is disclosed in copendingU.S. application Ser. No. 150,530, filed May 16, 1980, now U.S. Pat. No.4,351,771. Numerous analogs of spectinomycin, including correspondingprotected forms also among the present starting materials, are disclosedtherein. Therefore, U.S. application Ser. No. 150,530 is incorporatedherein by reference. Additional spectinomycin analogs are found incopending U.S. application Ser. No. 359,006, filed Mar. 17, 1982, nowU.S. Pat. No. 4,420,624; U.S. application Ser. No. 020,073, filed Mar.13, 1979, now U.S. Pat. No. 4,361,701; U.S. application Ser. No.312,035, filed Oct. 16, 1981, now U.S. Pat. No. 4,405,797; U.S.application Ser. No. 068,926, filed Aug. 23, 1979, now U.S. Pat. No.4,282,152; U.S. application Ser. No. 212,952, filed Dec. 4, 1980, nowU.S. Pat. No. 4,337,347; U.S. application No. Ser. No. 212,950, filedDec. 4, 1980, now U.S. Pat. No. 4,344,882; U.S. application No. Ser. No.212,943, filed Dec. 4, 1980, now U.S. Pat. No. 4,345,086; U.S.application Ser. No. 285,164, filed July 20, 1981, now U.S. Pat. No.4,380,651; U.S. application Ser. No. 285,165, filed July 20, 1981, nowU.S. Pat. No. 4,380,652; U.S. application Ser. No. 358,957, filed Mar.17, 1982, now U.S. Pat. No. 4,420,623; U.S. application Ser. No.359,723, filed Mar. 19, 1982, now abandoned, and U.S. application Ser.No. 314,261, filed Oct. 23, 1981 now abandoned. However, none of thesedisclosures appreciate the novel processes, novel intermediates, ornovel products of the present invention.

In general, preparation of Mannich bases are well known. Representativereferences of such preparations are March, "The Mannich Reaction",Advanced Organic Chemistry: Reactions, Mechanisms and Structure, pp.670-672, McGraw-Hill Book Company (1968); Flick, "The Use of a MannichBase as a Source of an Unsaturated Ketone for Condensations With anActive Methylene Compound", Organic Reactions, vol. 1, pp. 320-322, NewYork: John Wiley and Sons, Inc. (1942); Brewster et al. "AlkylationsWith Tertiary Amines", Organic Reactions, vol. 7, pp. 126-130, New York:John Wiley and Sons, Inc. (1953); Bergman et al. "Robinson'sModification of the Michael Condensation", Organic Reactions, vol. 10,pp. 222-223: New York, John Wiley and Sons, Inc. (1959). However, noneof these references makes obvious the present invention.

SUMMARY OF THE INVENTION

The present invention includes novel compounds having the formula

wherein R, R₁ and R₂ are as defined above;

The present invention also includes novel compounds having the formulaII wherein R₁, R₂, and R are as defined above.

Finally, the present invention includes novel compounds having theformula V

wherein R₃ and R₄ are the same and are hydrogen or blocking groups;

R is defined as above, and

--T is selected from the group consisting of --OY, --CHR₅ C(O)R₆, --NH₂,--NHR₅, --NR₅ R₆, --NO₂, --NHNH₂, --N₃, --NR₅ SO₂ R₆, --SH, --SR₅,--S--S--, --CH[C(O)R₅ ][C(O)R₆ ], --C.tbd.N, --CH(CN)R₅, --CH[C(O)R₅][C(O)OR₆ ], --CH[C(O)OR₅ ][C(O)OR₆ ], --CR₅ R₆ NO₂, --CR₅ R₆ SO₂ R₇,--CR₅ C(N)C(N)

wherein Y is aryl, --CR₅ ═CHR₆ or --N═O, and

R₅, R₆, and R₇ are the same or different and are lower alkyl or aryl andbiologically acceptable salts thereof.

In the foregoing designation of variables alkyl of C₁ to C₂₀ meansmethyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl,decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl,heptadecyl, octadecyl, nonadecyl, eicosyl, and isomers thereof. Thisdesignation is meant to include the preferred alkyl group, straight,cyclic or branch chain system in which the longest extension of thecyclic or branch chain system contains from 1 to 5 carbon atoms,inclusive.

Lower alkyl, --(CH₂)n-- wherein n is an integer of zero through four,and alkyl as is found in the terms lower haloalkyl or lower aminoalkylmeans an alkyl of from one through four carbon atoms, for example,methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, andtert-butyl.

Halo means fluoro, chloro, bromo or iodo.

Lower alkenyl means ethylidene, propylidene, butylidene, pentylidene,hexylidene, heptylidene, octylidene and the isomeric forms thereof.

Lower alkynyl means ethynyl, propynyl, butynyl, pentynyl, hexynyl,heptynyl, octynyl and the isomeric forms thereof.

Acyl means formyl, acetyl, propionyl, butyryl, pentanoyl and isomericforms thereof.

Blocking groups mean aralkoxycarbonyl, halogenated alkoxy carbonyl, andalkoxy carbonyl, such as, for example, 4-methoxy benzyloxycarbonyl,benzyloxycarbonyl, isopropoxycarbonyl, isobutyloxycarbonyl,cyclopentyloxycarbonyl, cyclohexyloxycarbonyl,β,β,β-trichloroethoxycarbonyl, or tertiary-butoxycarbonyl, preferablyeither benzoxycarbonyl or tertiary butoxycarbonyl.

Another aspect of the present invention is a process for preparingcompounds having the formula IV

wherein R₃ and R₄ are the same and are hydrogen or blocking groups;

R is as defined above, and

Q is a nucleophile which comprises

(1) contacting the compound III' with N,N-dimethyl(methylene)ammoniumchloride, that is, CH₂ ═N(CH₃)₂ Cl and trifluoroacetic acid (TFA) inacetonitrile (CH₃ CN) to obtain compound III";

(2) enolization of compound III" to obtain enone I;

(3) contacting the product of steps 1 or 2 with a nucleophile andtriethylamine (Et₃ N) to obtain the compound IVa and then, if desired;

(4) further treating IVa to obtain IVb

A preferred embodiment of the nucleophile --Q comprises --OZ,--(CH₂)_(b) OZ, --CHR₅ C(O)R₆, --NH₂, --NHR₅, --NR₅ R₆, --NO₂, --NHNH₂,--N₃, --NR₅ SO₂ R₆, --SH, --SR₅, --S--S--, --C.tbd.CR₅, --CH[C(O)R₅][C(O)R₆ ], --C.tbd.N, --CHCNR₅, --CH[C(O)R₅ ][C(O)OR₆ ], --CH[C(O)OR₅][C(O)OR₆ ], --CR₅ R₆ NO₂, --CR₅ R₆ SO₂ R₇, --CR₅ (CN)₂, hydrogen, loweralkyl, lower alkenyl, lower haloalkyl, lower aminoalkyl, or loweralkynyl

wherein Z is hydrogen, aryl, --N═O, lower alkyl, lower alkenyl, benzylor acyl;

b is an integer of one through eight;

R₁, R₂, R and R₃ through R₇ are all as defined above (see Scheme A).

The above process may be carried out without separately conducting steps1 and 2 above which identifies compound III" and the enone I. Thus, theproduct of step 1 may be contacted with the nucleophile and Et₃ N toobtain the desired compound IVa (see Scheme D).

The present invention includes an alternate process for the preparationof compounds having the formula IVa

wherein R₁, R₂, R, and Q are all as defined above which comprises

(1) contacting the compound III' withN,N-dimethylformamide-ditertiary-butyl acetal and trifluoroacetic acidin dimethylformamide (TFA/DMF) to obtain compounds having the formulaII;

(2) contacting the product of step 1 with sodium cyanoborohydride(NaCNBH₃) in methanol at about pH 4 to provide compound III";

(3) enolization of compound III" to obtain compound I;

(4) contacting the product of steps 2 or 3 with a nucleophile in thepresence of triethylamine (Et₃ N) to obtain compound IVa (see Scheme B).

Finally, the present invention includes the processes for thepreparation of compounds having the formula IVa

wherein R₁, R₂, R, and Q are all as defined above which comprises

(1) contacting the compound III' withN,N-dimethylformamide-ditertiary-butyl acetal and trifluoro acetic acidand dimethylformamide to obtain the compound II;

(2) contacting the product of step 1 with sodium cyanoborohydride inmethanol at pH 4 and treating the mixture with the nucleophile andtriethylamine to give the compound IVa (see Scheme C).

The compound IVa prepared in the final step of each of the abovealternate processes may also be further deprotected to obtain thedesired spectinomycin analogs IVb. Deprotection may be accomplished byknown methods in the art or by contacting the compound IVa withthioanisole (phenyl SCH₃ /TFA in neat trifluoroacetic acid). See step 4in Scheme A, step 5 in Scheme B, and step 3 in Schemes C and D.

Spectinomycin analogs of this invention are meant to includebiologically acceptable acid addition salts.

Biologically acceptable acid addition salts of the invention compoundsand the compounds prepared by the invention processes can be made byneutralizing the compounds with an appropriate acid to below about pH7.0 and advantageously to about from pH 2 to about pH 6. Suitable acidsfor this purpose include tartaric, gluconic, lactic, hydrochloric,sulfuric, phosphoric, sulfamic, hydrobromic and the like. Acid salts ofspectinomycin analogs can be used for the same biological purposes asthe parent compound.

The term "nucleophile" is meant to be nonlimiting, and the skilledartisan can readily determine operable groups from a broad range ofpossible reacting groups in the art known for the presence thereon of anunshared pair of electrons, and therefore, are included in the term"nucleophile." See March, Advanced Organic Chemistry: Reactions,Mechanisms, and Structure pp. 199-200 and pp. 287-290, McGraw-Hill BookCo. (1968) and Cram et al. Organic Chemistry, pp. 272-275, McGraw-HillBook Co. Preferred nucleophiles for use in the processes of thisinvention comprise HOM₁, R₅ OH, R₅ OM₁, R₅ CH═C(OM₁)R₆, NH₃, R₅ NH₂, R₅R₆ NH, NO₂ M₁, NH₂ NH₂, N₃ M₁, R₅ SO₂ NR₆ M₁, H₂ S, HSM₁, R₅ SH, S₂ M₂,R₅ M₁, R₅ C.tbd.CM₁, R₅ C(O)CHC(O)R₆ M₁ ←→R₅ C(OM₁)═CH(O)R₆ ←→R₅C(O)CH═C(OM₁)R₆, M₁ C.tbd.N, R₅ CHC.tbd.N, RSM₁, R₅ C(O)CHC(O)OR₆ M₁, R₅OC(O)CHC(O)OR₆ M₁, R₅ R₆ C(M₁)NO₂, R₅ R₆ C(M₁)SO₂ R₇ orHC(R₅)(C.tbd.N)(C.tbd.N), HR₈ or M₁ R₈

wherein R₅, R₆, and R₇ are the same or different and are lower alkyl oraryl; R₈ is lower alkyl, lower alkenyl, lower haloalkyl, loweraminoalkyl or lower alkynyl;

M₁ is a monovalent metal or ion, and

M₂ is a divalent metal.

Aryl with respect to the present invention includes phenyl and phenylsubstituted with, for example, 1 to 2 alkyl, alkoxy, or alkylthio suchas methylthio so that the named moiety is a substantially hydrocarbylmoiety.

The monovalent metal denoted M₁ herein is Na⁺, K⁺, or Li⁺.

The divalent metal denoted M₂ herein is Mg⁺⁺, Ca⁺⁺, or Ba⁺⁺.

DETAILED DESCRIPTION OF THE INVENTION

The functionalization of the compound III' withN,N-dimethyl-(methylene)ammonium chloride to obtain compound III" shownas step 1 in Scheme A occurs in a solution of acetonitrile or similarsolvents such as tetrahydrofuran, or 1,2-dimethoxyethane in the presenceof trifluoroacetic acid. The reaction is most efficaciously run at atemperature of 20° to 70° with molar ratios of compound III' toN,N-dimethyl(methylene)ammonium chloride to trifluoroacetic acid in theranges from 8:10:3 to 8:30:10. Preferably, reaction conditions are at atemperature of 35° C. to 45° C. using acetonitrile as a solvent with theratio of compound III' to N,N-dimethyl(methylene)ammonium chloride andtrifluoroacetic acid from 8:20:6 to 8:25:10.

A major competing side reaction at elevated temperatures occursfollowing the beta elimination shown in step 2 of Scheme A and step 3 ofScheme B to give the enone I. The side reaction is subsequentlydimerized to a product having formula VII and formula VIII shown in step3 of Scheme A and step 4 shown in Scheme B. However, the product III"can be obtained free of any residual starting material, enone or dimerby simply partitioning of the crude product mixture between ethylacetate and dilute aqueous acid, such as, HCl, H₂ SO₄, or HBr. Theaqueous layer is lyophilized to give the compound III" which isextremely hygroscopic but stable at 25° C.

Generally, the functionalization of N,N'-protected compound III' toobtain compound III" may also be accomplished by the preparation of anenamine II as shown in steps 1 of Scheme B and Scheme C. Thisfunctionalization is accomplished by usingN,N-dimethylformamide-ditertiary-butylacetal and trifluoroacetic acid indimethylformamide as shown in step 1 of Scheme B. A solution of III' indimethylformamide is cooled to about 0° C. to which is added theN,N-dimethylformamide-di-tert-butylacetal and trifluoroacetic acid. Thereaction mixture is allowed to warm and reaction proceeds smoothly atfrom 15° C. to 35° C., preferably about 25° C. Other reagents such asN,Ndimethylformamide-dimethylacetal in aprotic solvents such asacetonitrile or dimethylformamide may be used as shown in step 1 ofScheme C. However, N,N-dimethylformamide-dimethylacetal gives only lowyields of the desired enamines. Thus, care must be taken to avoidobtaining the rearranged compound N,N'protected actinospectinoic acidmethylester or analogs thereof having the formula VI. The enamine II isthen reduced with sodium cyanoborohydride in methanol at a pH from 3.5to 6, preferably about pH 4 to obtain theN,N'-diprotected-4'-(dimethylaminomethyl)compound as a hydrochloridesalt III" as shown in step 2 of both Scheme B and Scheme C. Maintenanceof the acidic pH during reduction is critical for obtaining the productIII" of this step. It is extracted with dilute base, such as NaHCO₃.

On the other hand, it appears that the trifluoroacetic acid catalyzesthe functionalization reaction of step 1 in each of the schemes shown byenolyzing the carbonyl of the protected starting material III'. It isbelieved that such enolization occurs to the carbonyl containingcompound III", thus providing a route for β elimination and subsequentreaction with the nucleophile shown in each of the Schemes A through D.Although, the enone I wherein R is methyl of the following examples isnot isolated, evidence of its existence may be found in fractionsobtained from chromatography of the reaction mixtures described abovefor obtaining compounds III". Attempts to concentrate these fractionsproduce a dimer having the formulae VII and VIII shown in Schemes A andB. The dimeric structure of compound VII and compound VIII, homogeneousby TLC, is evidenced by the number of signals in the CMR and offresonance decoupling experiments.

Since the enone, free of solvent, is not isolated in the presentinvention it is shown in brackets in Scheme A and Scheme B. Thus, the pHof the reaction product mixture obtained in step 1 of Scheme A and step2 of Scheme B described above is raised from pH of 7 to pH of 9,preferably from between pH 7.5 to pH 8.0 with triethylamine in thepresence of a nucleophile to obtain the product IVa. The reaction withthe nucleophile is conducted at from -50° C. to 40° C., preferably 25°C. for about 3 hours, preferably from two to four hours.

A one pot procedure as shown by step 2 of Scheme C may be moreefficacious. In the one pot reaction a solution of the enamine IIobtained by the above described procedure is dissolved in a loweralcohol, such as ethanol, propanol, or methanol at a pH of from 3 to 5,preferably about pH 4. A solution of sodium cyanoborohydride in methanolis added to the enamine solution while maintaining the pH at about fourwith methanolic hydrogen chloride. When the reaction mixture containsonly the amine III" and enone I as shown by thin layer chromatographythe reaction is complete. The nucleophile and triethylamine are addedand the reaction is stirred for 3 hours at 25° C.

Likewise, a one pot reaction as shown in Scheme D may be moreefficacious to accomplish the reation as previously described as stepsof Scheme A. In this reaction the conditions are as described for steps1 and 3 of Scheme A above with the added advantage that intermediatesIII" and I are not necessarily isolated or identified.

Finally, in step 4 of Scheme A, step 5 of Scheme B, step 3 of Scheme C,and step 3 of Scheme D the compound IVa is deprotected to obtain thedesired active compound IVb. Deprotection means removing the blockinggroups from the nitrogen in the actinamine ring of IVa and replacingthem with hydrogen. The particular conditions of the deprotection stepdepend upon the particular oxy groups, that is, group R₁ or R₂ thatblock the nitrogen on the actinamine ring. Further, the R substituent aswell as the nucleophile added by the present invention process toprepare compounds IVa determine the particular conditions for thedeprotection step described here. Many conventional methods may be usedas previously described in U.S. application Ser. No. 020,073, filed Nov.23, 1979, (Case 3589) if appropriate. However, since the nucleophile maypoison the catalyst in attempts to deprotect the compound IVa thendeprotection may be accomplished using thioanisole in neattrifluoroacetic acid. This is preferred.

Preferred compounds of the present invention are those wherein R is analkyl group, straight, cyclic or branch chain system, in which thelongest extension of the cyclic or branch chain system contains from 1to 5 carbon atoms, inclusive. Such an R substituent is as disclosed inU.S. application Ser. No. 359,723, filed Mar. 19, 1982 (Case 4034) and,therefore, is incorporated by reference. Most preferred are thosewherein R is n-butyl, n-pentyl or n-hexyl.

The following examples are indicative of the present invention and arenot to be construed as limiting. Those skilled in the art will readilyrecognize the appropriate variations from the procedure both as tovariations on the starting material III' as well as reaction conditionsand techniques. These examples indicate the best mode presently known tothe inventor. In each ml is milliliters; g is grams; mMol is millimole;C. is degrees centigrade; ppm is parts per million.

EXAMPLE I

Preparation ofN,N'-dicarbobenzyloxy-4'-(dimethyl-aminomethyl)-spectinomycin (III"wherein R₁ and R₂ are the same and are carbobenzyloxy, and R is methyl).See Scheme A step 1.

A solution of distilled acetyl chloride (1.50 ml, 1.58 g, 20.0 mMol) inether (5 ml) is added carefully to a solution ofN,N,N',N'-tetramethyldiaminomethane (2.70 ml, 2.02 g, 19.8 mMol) inether (50 ml). At 25° C. under N₂ atmosphere CH₂ ═N(CH₃)Cl precipitatesout immediately as a white solid. After 30 minutes the solvent isremoved with a filter stick, and the solid is washed two times with 25milliliters of ether. A CMR of a solid prepared by this procedure showed(d₃ -acetonitrile) 79.4, 38.8 ppm.

To this pre-formed reagent is added N,N'-dicarbobenzyloxyspectinomycin(III' wherein R₁ and R₂ are carbobenzyloxy, and R is methyl) (5.0 g, 8.3mMol), acetonitrile (25 ml), and trifluoroacetic acid (0.5 ml, 0.74 g,6.5 mMol). The substrate and reagent are stirred at 25° C. until asolution is effected. The reaction temperature is then raised to 40° C.and kept at 40° C. for a period of 72 hours. The reaction isconcentrated at 45° C. and the crude solid product is redissolved in 150ml of 0.05N hydrogen chloride and 150 ml of ethyl acetate. The layersare separated, and the organic layer is reextracted with 75 ml of theacid. Th aqueous fractions are combined, and the excess acid anddissolved ethyl acetate are removed on the rotary evaporator at 45°using a nitrogen stream. The sample is frozen and lyophilized to give 6g of the crude productN,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-spectinomycin (III"wherein R₁ and R₂ are carbobenzyloxy, and R is methyl). It is kept undernitrogen in a dry box.

The product prepared by the above description shows the following CMR(d₃ -acetonitrile) 157.7, 138.5, 129.7, 129.0, 128.8, 95.4, 91.4, 74.5,74.0, 69.6, 67.9, 66.5, 66.2, 60.8, 57.6, 51.2, 47.0, 35.5, 32.5, 18.2ppm.

Following the procedure in Example I, but substituting the appropriateN,N'-dicarbobenzyloxy spectinomycin (III' wherein R₁ and R₂ arecarbobenzyloxy, and R is selected from the group as defined above) thefollowing N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-spectinomycinderivative III" can be prepared.

1.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-5'-demethylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

2.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-hydroxyspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is hydroxymethyl);

3. N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-chlorospectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is chloromethyl);

4. N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-bromospectinomycin)(III" wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl);

5.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-n-butylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is pentyl);

6.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-acetoxyspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is acetoxymethyl);

7.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-phenylmethoxyspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxymethyl);

8.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-5'-demethyl-5'-phenylmethoxyspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxy);

9.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-5'-demethyl-5'-methoxyspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

10.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-methylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl);

11.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-methoxyspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is methoxymethyl);

12. N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-aminospectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is aminomethyl);

13. N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-ethylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is propyl);

14.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-n-pentylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is n-hexyl);

15.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-5'-dimethyl-5'-methylethynyldemethylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is methylethynyl);

16.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-n-propylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl);

17.N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-6'-ethenyloxymethylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is ethenyloxyethyl);

EXAMPLE II

Preparation of 4'-Enamine,N,N'-dicarbobenzyloxy-4'-dimethylaminomethylenespectinomycin (II whereinR₁ and R₂ are carbobenzyloxy, and R is methyl). See Scheme B step 1 orScheme C step 1.

To a solution of N,N'-dicarbobenzyloxyspectinomycin (III' wherein R₁ andR₂ are carbobenzyloxy, and R is methyl) (5.0 g, 8.33 mMol) indimethylformamide (10 ml) cooled to 0° C. is addedN,N-dimethylformamide-di-tert-butyl-acetal (9 ml, 7.6 g, 37.6 mMol) andtrifluoroacetic acid (0.9 ml, 1.35 g, 11.8 mMol). The reaction is cooledan additional 15 minutes then allowed to warm to 25° C. The reaction isnearly complete after seven hours. It is stoppered and stored at -5° C.for two days. The solution is then concentrated at 45° C. to an oilysolid. The crude product is then chromatographed on 250 g of silica gel(E. Merck Silica Gel 60) using medium pressure and a gradient elutionfrom 1:9 to 1:1 acetone:chloroform. The4'-enamine,N-N'-dicarbobenzyloxy-4'-dimethylaminomethylenespectinomycin,prepared by this method (II, wherein R₁ and R₂ are carbobenzyloxy, and Ris methyl) (2.1 g, 38.7%) was isolated as an orange colored solid andcharacterized as follows:

High Resolution Mass Spec.: (Tri-TMS) calcd for C₄₂ H₆₅ N₃ O₁₁ Si₃=871.3927, Found: 871.3955. CMR (d₆ -Acetone): 186.7, 157 (m), 154.6,138.1, 128.4, 129.1, 102.9, 95.0, 89.5, 74.7, 74.1, 69.5, 67.2, 66.5,65.5, 60 (m), 57.0 (m), 31.6, 25.2 ppm. PMR (CDCl₃): δ 1.4 (CH₃, d), 3.0(NCH₃, s), 3.7-4.8 (m), 5.0 (--CH₂ --o, s), 7.3 (ArH, s).

IR (Mull) 3400 s (OH), 1690 s (C=O), 1585, 1555 s, 1495 (C=C), 1420,1345, 1300, 1165, 1080, 1060, 770, 740, 695 cm⁻¹.

CD (MeOH) [θ]₃₄₃ nm^(max) =-9,200±1,100; [θ]₂₇₄ nm^(max) =+4,600±1,100

CD (CH₃ CN) [θ]₃₄₆ nm^(max) =-11,500±1,000

[α]D (CHCl₃, C=0.483)=-9°; UV (CH₃ CN) 204 sh (ε=22,750) 334 (ε=13,950).

Following the procedure in Example II but substituting the appropriatelysubstituted N,N'-dicarbobenzyloxyspectinomycin (III' wherein R₁ and R₂are carbobenzyloxy, and R is selected from the group defined above) thefollowing novel 4'-enamine is prepared.

1.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-5'-demethylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

2.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-hydroxyspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is hydroxymethyl);

3.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-chlorospectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is chloromethyl);

4. N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-bromospectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl);

5.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-n-butylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is pentyl);

6.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-acetoxyspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is acetoxymethyl);

7.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-phenylmethoxyspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxymethyl);

8.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-5'-demethyl-5'-phenylmethoxyspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxy);

9.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-5'-demethyl-5'-methoxyspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

10.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-methylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl);

11.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-methoxyspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is methoxymethyl);

12.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-aminospectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is aminomethyl);

13.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-ethylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is propyl);

14.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-n-pentylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is n-hexyl);

15.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-5'-demethyl-5'-methylethynylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is methylethynyl);

16.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-n-propylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl);

17.N,N'-dicarbobenzyloxy-4'-dimethylaminomethylene-6'-ethenyloxymethylspectinomycin(II wherein R₁ and R₂ are carbobenzyloxy, and R is ethenyloxyethyl);

EXAMPLE III

Preparation ofN,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-spectinomycin hydrogenchloride (III" wherein R₁ and R₂ are carbobenzyloxy, and R is methyl),4'-enone (I, wherein R₁ and R₂ are carbobenzyloxy, and R is methyl) anddimer (VII and VIII wherein R₁ and R₂ are carbobenzyloxy). See Scheme Bsteps 2, 3, and 4.

To a solution of the 4'-enamine,N,N'-dicarbobenzyloxy-4'-dimethylaminomethylenespectinomycin, fromExample II above (414 mg, 0.63 mMol) in methanol (5 ml) at pH 4 is addedsodium cyanoborohydride (13.2 mg, 0.21 mMol) over a period of 10minutes. The pH of the solution is readjusted after each addition ofhydride with methanolic hydrogen chloride using methyl orange as anindicator. The reaction is stirred at 25° C. for two hours thenconcentrated to give the crude product mixture. Two major product spotsare observed by TLC in 1:9 MeOH:CHCl₃, the 4'-enone,N,N'-dicarbobenzyloxy-4'-methylenespectinomycin, (I wherein R₁ and R₂are carbobenzyloxy, and R is methyl) is less polar (Rf 0.6) than thestarting material 4'-enamine,N,N'-dicarbobenzyloxy-4'-dimethylaminomethylenespectinomycin, (Rf 0.5)with intense UV and permanganate activity, and the amine,N,N'-di-carbobenzyloxy-4'-(dimethylaminomethyl)-spectinomycin hydrogenchloride (III" wherein R₁ and R₂ are carbobenzyloxy, and R is methyl)which remains at the origin. The crude product mixture is redissolved inchloroform (60 ml), extracted with 0.05M NaOH (10 ml) and water (20 ml)then dried over Na₂ SO₄ and concentrated to give 375 mg of the dimer(VII and VIII wherein R₁ and R₂ are carbobenzyloxy). The sample ischromatographed on silica gel 60 (40 g) using 2 percent MeOH:CHCl₃.Fractions containing the pure 4'-enone,N,N'-dicarbobenzyloxy-4'-methylenespectinomycin, (I wherein R₁ and R₂are carbobenzyloxy, and R is methyl) are pooled to give 100 mg ofmaterial. TLC of this pooled fraction shows that the 4'-enone isconverted in part to the more polar spot dimer. Almost completeconversion to the dimer was observed when a 4'-enone prepared by thismethod was allowed to stand in solution.

It showed a CMR (d₆ -acetone): 193.3, 157.8, 157.0, 143.1, 138.2, 129.2,128.4, 111.2, 97.6, 95.9, 93.8, 88.7, 82.7, 75.2, 74.5, 72.4, 71.2,67.2, 66.8, 65.1, 31.5, 22.5, 20.0, 17.7, 14.1 ppm.

Following the procedure in Example III but substituting the appropriateN,N'-dicarbobenzyloxy-4'-dimethylaminomethylenespectinomycin (II whereinR₁ and R₂ are carbobenzyloxy, and R is selected from the group asdefined above) the following 4'-enone is prepared.

1. N,N'-dicarbobenzyloxy-4'-methylene-5'-demethylspectinomycin (Iwherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

2. N,N'-dicarbobenzyloxy-4'-methylene-6'-hydroxyspectinomycin (I whereinR₁ and R₂ are carbobenzyloxy, and R is hydroxymethyl);

3. N,N'-dicarbobenzyloxy-4'-methylene-6'-chlorospectinomycin (I whereinR₁ and R₂ are carbobenzyloxy, and R is chloromethyl);

4. N,N'-dicarbobenzyloxy-4'-methylene-6'-bromospectinomycin (I whereinR₁ and R₂ are carbobenzyloxy, and R is bromomethyl);

5. N,N'-dicarbobenzyloxy-4'-methylene-6'-n-butylspectinomycin (I whereinR₁ and R₂ are carbobenzyloxy, and R is pentyl);

6. N,N'-dicarbobenzyloxy-4'-methylene-6'-acetoxyspectinomycin (I whereinR₁ and R₂ are carbobenzyloxy, and R is acetoxymethyl);

7. N,N'-dicarbobenzyloxy-4'-methylene-6'-phenylmethoxyspectinomycin (Iwherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxymethyl);

8.N,N'-dicarbobenzyloxy-4'-methylene-5'-demethyl-5'-phenylmethoxyspectinomycin(I wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxy);

9.N,N'-dicarbobenzyloxy-4'-methylene-5'-demethyl-5'-methoxyspectinomycin(I wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

10.N,N'-dicarbobenzyloxy-4'-methylene-5'-demethyl-6'-methylspectinomycin (Iwherein R₁ and R₂ are carbobenzyloxy, and R is ethyl);

11.N,N'-dicarbobenzyloxy-4'-methylene-5'-demethyl-6'-methoxyspectinomycin(I wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

12. N,N'-dicarbobenzyloxy-4'-methylene-6'-aminospectinomycin (I whereinR₁ and R₂ are carbobenzyloxy, and R is aminomethyl);

13. N,N'-dicarbobenzyloxy-4'-methylene-6'-ethylspectinomycin (I whereinR₁ and R₂ are carbobenzyloxy, and R is propyl);

14. N,N'-dicarbobenzyloxy-4'-methylene-6'-n-pentylspectinomycin (Iwherein R₁ and R₂ are carbobenzyloxy, and R is n-hexyl);

15.N,N'-dicarbobenzyloxy-4'-methylene-5'-demethyl-5'-methylethynylspectinomycin-4'-ylenone (I wherein R₁ and R₂ are carbobenzyloxy, and R is methylethynyl);

16. N,N'-dicarbobenzyloxy-4'-methylene-6'-n-propylspectinomycin (Iwherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl);

17. N,N'-dicarbobenzyloxy-4'-methylene-6'-ethenyloxymethylspectinomycin(I wherein R₁ and R₂ are carbobenzyloxy, and R is ethenyloxyethyl);

Likewise the procedure in Example III having the appropriatelysubstituted enamine for preparing the 4'-enone is then also converted tothe following novel dimers:

1. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen;

2. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen;

3. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R ishydroxymethyl;

4. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R ishydroxymethyl;

5. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R ischloromethyl;

6. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R ischloromethyl;

7. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R isbromomethyl;

8. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R isbromomethyl;

9. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R is pentyl;

10. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R is pentyl;

11. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R isacetoxymethyl;

12. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R isacetoxymethyl;

13. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxymethyl;

14. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxymethyl;

15. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxy;

16. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxy;

17. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy;

18. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy;

19. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl;

20. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl;

21. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R ismethoxymethyl;

22. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R ismethoxymethyl;

23. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R isaminomethyl;

24. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R isaminomethyl;

25. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R is propyl;

26. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R is propyl;

27. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R is n-pentyl;

28. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R isn-pentyl;

29. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R ismethylethynyl;

30. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R ismethylethynyl;

31. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl;

32. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl;

33. Formula VII wherein R₁ and R₂ are carbobenzyloxy, and R isethenyloxyethyl;

34. Formula VIII wherein R₁ and R₂ are carbobenzyloxy, and R isethenyloxyethyl.

EXAMPLE IV

Preparation ofN,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-spectinomycin (IVa whereinR₁ and R₂ are carbobenzyloxy; R is methyl, and T is --S--CH₂ CH₂ CH₂CH₂). See Scheme B steps 2, 3, and 4a or Scheme C step 2.

To a solution of the 4'-enamine,N,N'-dicarbobenzyloxy-4'-dimethylaminomethylenespectinomycin, fromExample II above (500 mg, 0.76 mMol) dissolved in methanol (7.5 ml) atpH 4 is added a solution of sodium cyanoborohydride (160 mg, 0.25 mMol)in methanol (2.5 ml) over a period of ten minutes. The pH is maintainedat pH 4 by methanolic hydrogen chloride. TLC after one hour shows onlythe N,N'dicarbobenzyloxy-4'-(dimethylaminomethyl)-spectinomycin (III"wherein R₁ and R₂ are carbobenzyloxy, and R is methyl) and 4'-enone,N,N'-dicarbobenzyloxy-4'-methylenespectinomycin, (as is identified inExample III above). Butanethiol (0.5 ml, ˜0.4 g, ˜4.4 mMol) andtriethylamine (0.06 ml) are added, and the reaction is stirred for twomore hours at 25° C. The reaction is concentrated and chromatographed onsilica gel 60 (45 g) using an acetone:chloroform gradient elution(1:9-2:8). 120 mg, a yield of 22 percent by weight of theN,N'-dicarbobenzyloxy-4'(n-butylthiomethyl)-spectinomycin (IVa whereinR₁ and R₂ are carbobenzyloxy, R is methyl, and Q is --S--CH₂ CH₂ CH₂CH₂). This compound is identified by comparison to the product in thefollowing Example V.

Following the procedure in Example IV but substituting the appropriate4'-enamine, N,N'-dicarbobenzyloxy-4'-dimethylaminomethylspectinomycin,as prepared in Example II above (II wherein R₁ and R₂ arecarbobenzyloxy, and R is selected from the group as defined above) andfurther substituting the appropriate nucleophile for butanethiol thefollowing novel N,N'-di-carbobenzyloxy-4'-(substituted) spectinomycinderivative is prepared.

1. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

2. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-hydroxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydroxymethyl);

3. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-chlorospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is chloromethyl);

4. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-bromospectinomycin)(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl);

5. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-n-butylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is pentyl);

6. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-acetoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is acetoxymethyl);

7.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-phenylmethoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxymethyl);

8.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-5'-phenylmethoxydesmethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxy);

9.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-5'-methoxy-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

10. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-methylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl);

11. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-methoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxymethyl);

12. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-aminospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is aminomethyl);

13. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-ethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is propyl);

14.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-n-pentylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-hexyl);

15.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-5'-methylethynyl-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methylethynyl);

16.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-n-propylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl);

17.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-ethenyloxymethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is ethenyloxyethyl);

18. N,N'-dicarbobenzyloxy-4'-(nitrosooxymethyl)-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

19.N,N'-dicarbobenzyloxy-4'-[2-(N-phenyl-N-propylamino)ethyl]-6'-bromospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl);

20. N,N'-dicarbobenzyloxy-4'-(nitromethyl)-6'-n-butylspectinomycin (IVawherein R₁ and R₂ are carbobenzyloxy, and R is pentyl);

21. N,N'-dicarbobenzyloxy-4'-(hydrazinomethyl)-6'-acetoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is acetoxymethyl);

22. N,N'-dicarbobenzyloxy-4'-(azidomethyl)-6'-phenylmethoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxymethyl);

23.N,N'-dicarbobenzyloxy-4'-[2-[[(methylsulfonyl)methyl]amino]-ethyl]-5'demethyl-5'-phenylmethoxydemethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxy);

24.N,N'-dicarbobenzyloxy-4'-(mercaptomethyl)-5'-demethyl-5'-methoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

25. N,N'-dicarbobenzyloxy-4'-[(methylthio)methyl]-6'-methylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl);

26. Bis-[N,N'-dicarbobenzyloxy-6'-methoxyspectinomycin-4'-yl]-disulfide(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxymethyl);

27.N,N'-dicarbobenzyloxy-4'-(2-acetyl-2-propionylethyl-6'-n-pentylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-hexyl);

28.N,N'-dicarbobenzyloxy-4'-(cyanomethyl)-5'-demethyl-5'-methylethynylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methylethynyl);

29. N,N'-dicarbobenzyloxy-4'-(3-cyanopropyl)-6'-n-propylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl);

30.N,N'-dicarbobenzyloxy-4'-[2-acetyl-2-[(ethenyloxy)carbonyl]]ethyl-6'-(2-ethenyloxymethylspectinomycin)(III" wherein R₁ and R₂ are carbobenzyloxy, and R is ethenyloxyethyl);

31.N,N'-dicarbobenzyloxy-4'-[2,2-bis(ethoxycarbonyl)-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

32.N,N'-dicarbobenzyloxy-4'-(2,2-dimethyl-2-nitroethyl)-6'-hydroxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydroxymethyl);

33.N,N'-dicarbobenzyloxy-4'-[2,2-diethyl-2-(methylsulfonyl)-ethyl]-6'-chlorospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is chloromethyl);

34.N,N'-dicarbobenzyloxy-4'-(2,2-cyano-2-isobutylethyl)-6'-bromospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl).

EXAMPLE V

Preparation ofN,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-spectinomycin (IVa whereinR₁ and R₂ are carbobenzyloxy; R is methyl, and Q is --S--CH₂ CH₂ CH₂CH₂). See Scheme D steps 1 and 2.

N,N-dimethyl(methylene)ammonium chloride

A solution of distilled acetyl chloride (1.50 ml, 1.58 g, 20.0 mMol) inether (5 ml) is added carefully to a solution ofN,N,N',N'-tetramethyldiamino-methane (2.70 ml, 2.02 g, 19.8 mMol) inether (50 ml). At 25° C. under N₂ atmosphere the product precipitatesout immediately as a white solid. After 30 minutes, the solvent isremoved with a filter stick, and the solid is washed two times with 25milliliters of ether. CMR (d₃ -acetonitrile). 79.4, 38.8 ppm.

N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-spectinomycin hydrogenchloride (III" wherein R₁ and R₂ are carbobenzyloxy, and R is methyl)

To the pre-formed reagent described above is addedN,N'-dicarbobenzyloxyspectinomycin (III" wherein R₁ and R₂ arecarbobenzyloxy, and R is methyl) (5.0 g, 8.3 mMol), acetonitrile (25 ml)and trifluoroacetic acid (0.5 ml, 0.74 g, 6.5 mMol). The substrate andreagent are stirred at 25° C. until a solution is effected. The reactionis then refluxed for 2.5 hours and cooled to 25° C. before the additionof n-butane thiol.

N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-spectinomycin (IVa whereinR₁ and R₂ are carbobenzyloxy, and R is methyl)

To the above solution of theN,N'-dicarbobenzyloxy-4'-dimethylaminomethyl)-spectinomycin hydrogenchloride (III" wherein R₁ and R₂ are carbobenzyloxy, and R is methyl)and the 4'-enone, N,N'-dicarbobenzyloxy-4'-methylenespectinomycin, (Iwherein R₁ and R₂ are carbobenzyloxy, and R is methyl) is addedtert-butanol (15 ml) and n-butanethiol (4 ml, 3.2 g, 35.5 mMol). The pHof the reaction mixture is adjusted to between pH 7.5-8.0 withtriethylamine. The reaction is stirred at 25° C. for 3.5 hours thenplaced in the freezer over night. The solution is concentrated to an oiland chromatographed on silica gel 60 with an acetone:chloroform gradient(1:9→2:8). 3.15 g of theN,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-spectinomycin (IVa whereinR₁ and R₂ are carbobenzyloxy; R is methyl, and T is --S--CH₂ CH₂ CH₂CH₂). An oil prepared as described above showed the followingcharacteristics.

Mass spec: (mixture of tri- and tetra-TMS):

    3×TMS 918 M+, 903

    4×TMS 990 M+, 975, 901.

CMR (d₆ -acetone): 202.2, 157, 138.0, 129.1, 128.3, 97.2, 92.2, 75.0,74, 71.4, 67.2, 66.4, 65.7, 60.5, 57.4, 54.1, 33.7, 32.2, 31.4, 26.9,22.3, 20.1, 13.8 ppm.

PMR (CDCl₃) 7.26 (φ, 10H, s), 5.06 (OCH₂, 4H, s), 3.34-4.65 (CHOH, 11H,m), 3.04 (NCH₃, 6H, d), 2.71 (CH₂ --S, 2H, d), 2.53 (S--CH₂, 2H, t),1.44 (CH₃, 3H, d), 1.65-1.25 (CH₂, 5H, m), 0.89 (CH₃, 3H, t).

[α]_(D) =+3°.

I.R. 3400, 1685, 1585, 1500, 1545, 1345, 1165, 1115, 1080, 1060, 1030,770, 735, 695.

Following the procedure in Example V but substituting the appropriateanalog of N,N'-dicarbobenzyloxy-4'-(dimethylaminomethyl)-spectinomycinhydrogen chloride (III" wherein R₁ and R₂ are carbobenzyloxy, and R isselected from the group as defined above), and again furthersubstituting the appropriate nucleophile as defined above is substitutedfor butanethiol the followingN,N'-dicarbobenzyloxy-4'-(substituted)-spectinomycin derivative isprepared.

1. N,N'-dicarbobenzyloxy-4'-(n-butythiomethyl)-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

2. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-hydroxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydroxymethyl);

3. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-chlorospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is chloromethyl);

4. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-bromospectinomycin)(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl);

5. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-n-butylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is pentyl);

6. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-acetoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is acetoxymethyl);

7.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-phenylmethoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxymethyl);

8.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-5'-demethyl-5'-phenylmethoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxy);

9.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-5'-demethyl-5'-methoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

10. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-methylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl);

11. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-methoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxymethyl);

12. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-aminospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is aminomethyl);

13. N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-ethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is propyl);

14.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-n-pentylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-hexyl);

15.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-5'-demethyl-5'-methylethynylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methylethynyl);

16.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-n-propylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl);

17.N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-6'-ethenyloxymethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is ethenyloxyethyl);

18. N,N'-dicarbobenzyloxy-4'-(nitrosooxymethyl)-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

19.N,N'-dicarbobenzyloxy-4'-[2-(N-phenyl-N-propylamino)-ethyl]-6'-bromospectinomycin)(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl);

20. N,N'-dicarbobenzyloxy-4'-(nitromethyl)-6'-n-butylspectinomycin (IVawherein R₁ and R₂ are carbobenzyloxy, and R is pentyl);

21. N,N'-dicarbobenzyloxy-4'-(hydrazinomethyl)-6'-acetoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is acetoxymethyl);

22. N,N'-dicarbobenzyloxy-4'-(azidomethyl)-6'-phenylmethoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R isphenylmethoxymethyl);

23.N,N'-dicarbobenzyloxy-4'-[2-[[(methylsulfonyl)methyl]amino]-ethyl]-5'-phenylmethoxydesmethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is phenylmethoxy);

24.N,N'-dicarbobenzyloxy-4'-(mercaptomethyl)-5'-demethyl-5'-methoxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxy);

25. N,N'-dicarbobenzyloxy-4'-[(methylthio)methyl]-6'-methylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is ethyl);

26. Bis-[N,N'-dicarbobenzyloxy-6'-methoxyspectinomycin-4'-yl]-disulfide(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methoxymethyl);

27.N,N'-dicarbobenzyloxy-4'-(2-acetyl-2-propionylethyl)-6'-n-pentylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-hexyl);

28.N,N'-dicarbobenzyloxy-4'-(cyanomethyl)-5'-demethyl-5'-methylethynylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is methylethynyl);

29. N,N'-dicarbobenzyloxy-4'-(3-cyanopropyl)-6'-n-propylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is n-butyl);

30.N,N'-dicarbobenzyloxy-4'-(2-acetyl-2-[(ethenyloxy)carbonyl]-ethyl-6'-(2-ethenyl)oxymethylspectinomycin(III" wherein R₁ and R₂ are carbobenzyloxy, and R is 2-ethenyloxyethyl);

31.N,N'-dicarbobenzyloxy-4'-[(2,2-bis(ethoxycarbonyl)ethyl-5'-demethylspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydrogen);

32.N,N'-dicarbobenzyloxy-4'-(2,2-dimethyl-2-nitroethyl)-6'-hydroxyspectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is hydroxymethyl);

33.N,N'-dicarbobenzyloxy-4'-[2,2-diethyl-2-(methylsulfonyl)-ethyl]-6'-chlorospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is chloromethyl);

34.N,N'-dicarbobenzyloxy-4'-(2,2-cyano-2-isobutylethyl)-6'-bromospectinomycin(IVa wherein R₁ and R₂ are carbobenzyloxy, and R is bromomethyl).

EXAMPLE VI

Preparation of 4'-(n-butylthiomethyl)-spectinomycin (IVb wherein Q is--S--CH₂ CH₂ CH₂ CH₂). See deprotection steps of each Scheme A throughD.

The N,N'-dicarbobenzyloxy-4'-(n-butylthiomethyl)-spectinomycin (IVawherein R₁ and R₂ are carbobenzyloxy; R is methyl, and Q is --S--CH₂ CH₂CH₂ CH₃) (150 mg, 0.21 mMol) from Example V is stirred with thioanisole(1 ml) and distilled trifluoroacetic acid (4 ml) at 25° C. for four tofive hours. The reaction is concentrated, and the product isprecipitated with ether (20 ml) to a gum. The solid is washed with ether(25 ml) and redissolved in isopropanol (7 ml). Conversion to thedihydrochloride salt with 1N HCl in isopropanol gives a white solidprecipitate which is filtered to give 65 mg (61 percent) of4'-(n-butylthiomethyl)-spectinomycin (IVb wherein Q is --S--CH₂ --CH₂--CH₂ --CH₂). It was identified as follows:

CMR (D₂ O) 94.1, 93.0, 73.1, 70.8, 67.0, 66.7, 62.6, 60.5, 59.6, 49.8,33.2, 31.9, 31.3, 27.2, 22.4, 19.6, 14.1 ppm. High resolution mass spec.(tri-TMS) calcd for C₂₈ H₅₈ N₂ O₇ S₁ Si₃ =650.3272. Found:=650.3252.

Following the procedure in Example VI but substituting the appropriateN,N'-dicarbobenzyloxy-4'-(substituted)-spectinomycin (IVa wherein R₁ andR₂ are carbobenzyloxy; R and Q are selected from the group as definedabove), the following spectinomycin derivative is prepared.

1. 4'-(n-butylthiomethyl)-5'-demethylspectinomycin (IVb wherein R ishydrogen);

2. 4'-(n-butylthiomethyl)-6'-hydroxyspectinomycin (IVb wherein R ishydroxymethyl);

3. 4'-(n-butylthiomethyl)-6'-chlorospectinomycin (IVb wherein R ischloromethyl);

4. 4'-(n-butylthiomethyl)-6'-bromospectinomycin) (IVb wherein R isbromomethyl);

5. 4'-(n-butylthiomethyl)-6'-n-butylspectinomycin (IVb wherein R ispentyl);

6. 4'-(n-butylthiomethyl)-6'-acetoxyspectinomycin (IVb wherein R isacetoxymethyl);

7. 4'-(n-butylthiomethyl)-6'-phenylmethoxyspectinomycin (IVb wherein Ris phenylmethoxymethyl);

8. 4'-(n-butylthiomethyl)-5'-demethyl-5'-phenylmethoxyspectinomycin (IVbwherein R is phenylmethoxy);

9. 4'-(n-butylthiomethyl)-5'-demethyl-5'-methoxyspectinomycin (IVbwherein R is methoxy);

10. 4'-(n-butylthiomethyl)-6'-methylspectinomycin (IVb wherein R isethyl);

11. 4'-(n-butylthiomethyl)-6'-methoxyspectinomycin (IVb wherein R ismethoxymethyl);

12. 4'-(n-butylthiomethyl)-6'-aminospectinomycin (IVb wherein R isaminomethyl);

13. 4'-(n-butylthiomethyl)-6'-ethylspectinomycin (IVb wherein R ispropyl);

14. 4'-(n-butylthiomethyl)-6'-n-heptylspectinomycin (IVb wherein R isn-hexyl);

15. 4'-(n-butylthiomethyl)-5'-demethyl-5'-methylethynylspectinomycin(IVb wherein R is methylethynyl);

16. 4'-(n-butylthiomethyl)-6'-n-propylspectinomycin (IVb wherein R isn-butyl);

17. 4'-(n-butylthiomethyl)-6'-ethenyloxymethylspectinomycin (IVb whereinR is ethenyloxyethyl);

18. 4'-(oxynitrosooxymethyl)-5'-demethyl-5'-spectinomycin (IVb wherein Ris hydrogen);

19. 4'-(methylaminomethyl)-6'-hydroxyspectinomycin (IVb wherein R ishydroxymethyl);

20. 4'-[(ethylmethylamino)methyl]-6'-chlorospectinomycin (IVb wherein Ris chloromethyl);

21. 4'-[2'-(N-phenyl-N-propylamino)ethyl]-6'-bromospectinomycin (IVbwherein R is bromomethyl);

22. 4'-(nitromethyl)-6'-n-butylspectinomycin (IVb wherein R is pentyl);

23. 4'-(hydrazinomethyl)-6'-acetoxyspectinomycin (IVb wherein R isacetoxymethyl);

24. 4'-(azidomethyl)-6'-phenylmethoxyspectinomycin (IVb wherein R isphenylmethoxymethyl);

25.4'-[2-[[(methylsulfonyl)methyl]amino]ethyl]-5'-demethyl-5'-phenylmethoxyspectinomycin(IVb wherein R is phenylmethoxy);

26. 4'-(mercaptomethyl)-5'-demethyl-5'-methoxyspectinomycin (IVb whereinR is methoxy);

27. 4'-[(methylthio)methyl]-6'-methylspectinomycin (IVb wherein R isethyl);

28. Bis-(6'-methoxyspectinomycin-4'-yl)disulfide (IVb wherein R ismethoxymethyl);

29. 4'-(n-butyl)-6'-aminospectinomycin (IVb wherein R₁ and R₂ arecarbobenzyloxy, and R is aminomethyl);

30. 4'-[(methylethynyl)methyl]-6'-ethylspectinomycin (IVb wherein R ispropyl);

31. 4'-(2-acetyl-2-propionylethyl)-6'-n-pentylspectinomycin (IVb whereinR is n-hexyl);

32. 4'-(cyanomethyl)-5'-demethyl-5'-methylethynylspectinomycin (IVbwherein R is methylethynyl);

33. 4'-(3-cyanopropyl)-6'-n-propylspectinomycin (IVb wherein R isn-butyl);

34.4'-(2-acetyl-2-[(ethenyloxy)carbonyl]ethyl-6'-(2-ethenyl)oxymethylspectinomycin(IVb wherein R is ethenyloxyethyl);

35. 4'-[2,2-bis(ethoxycarbonyl)ethyl]-5'-demethylspectinomycin (IVbwherein R is hydrogen);

36. 4'-(2,2-dimethyl-2-nitroethyl)-6'-hydroxyspectinomycin (IVb whereinR is hydroxymethyl);

37. 4'-[2,2-diethyl-2-(methylsulfonyl)ethyl]-6'-chlorospectinomycin (IVbwherein R is chloromethyl);

38. 4'-(2,2-cyano-2-isobutylethyl)-6'-bromospectinomycin (IVb wherein Ris bromomethyl). ##STR1##

We claim:
 1. A compound of the formulawherein R₃ and R₄ are the same andare hydrogen or blocking groups; T is selected from the group consistingof --OY, --CHR₅ C(O)R₆, [--NH₂,] --NHR₅, --NR₅ R₆, --NO₂, --NHNH₂, --N₃,--NR₅ SO₂ R₆, --SH, --SR₅, --S--SM₂ ⁺, --CH[C(O)R₅ ]C(O)R₆ ], --C.tbd.N,--CHC(N)R₅, --CH[C(O)R₅ ][C(O)OR₆ ], --CH[C(O)OR₅ ][C(O)OR₆ ], --CR₅ R₆NO₂, --CR₅ R₆ SO₂ R₇, --CR₅ C(N)C(N), wherein Y is aryl or --CR₅ ═CHR₆or --N═O, and R₅, R₆, and R₇ are the same or different and are loweralkyl or aryl with the proviso that when T is --NHR₅, --NR₅ R₆ or --OCR₅═CHR₆, R₅ and R₆ are aryl; wherein R is hydrogen, alkyl of from C₁ toC₂₀, inclusive, lower alkenyl, lower haloalkyl, lower aminoalkyl, loweralkynyl or --(CH₂)_(n) --OX wherein X is selected from the groupconsisting of hydrogen, lower alkyl, lower alkenyl, benzyl, and acylwherein n is an integer of zero to four with the proviso that when n iszero --OX cannot be hydroxy and biologically acceptable salts thereof.2. A compound of claim 1 wherein R₃ and R₄ are a blocking group.
 3. Acompound of claim 1 wherein R₃ and R₄ are hydrogen.